Process for providing enhanced yield and simplified isolation of

Details Process for providing enhanced yield and simplified isolation of Process for providing enhanced yield and simplified isolation of

1991-05-20

1993-09-21

Naff, David M.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

4352533, 435874, C12N 920, C12N 120, C12N 100

Patent

active

052468488

ABSTRACT:
This invention is directed to a process for providing enhanced yield and simplified isolation of hydrophobic enzymes from a culture medium. The process comprises the steps of: (a) culturing a hydrophobic enzyme-secreting microorganism in an aqueous culture medium comprising a yield-enhancing effective amount of a nonionic surfactant having a cloud point of less than 40.degree. C. for a time sufficient to provide enhanced secretion of said enzyme from said microorganism, (b) removing said microorganism from said culture medium to provide a microorganism-free solution, (c) heating said microorganism-free solution to a temperature above said cloud point to cause phase separation of said culture medium into an aqueous phase, and a non-aqueous phase containing said nonionic surfactant and said yield-enhanced hydrophobic enzyme, and (d) isolating said non-aqueous phase containing said nonionic surfactant and said yield-enhanced hydrophobic enzyme.

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patent: 5063160 (1991-11-01), Holmes
Philippe Burgisser, Jean-Marie Matthieu, Phase Separation of Myelin Proteins in Triton X-114, 1989, 11:179-187.
Journal of Applied Biochemistry 7, 356-369 (1985) "Isolation of Human . . . Chromatography".
The Journal of Biological Chemistry, vol. 256 No. 4, Issue of Feb. 25, pp. 1604-1607, 1981, vol. 259, No. 23, Issue of Dec. 10, pp. 14922-14927, 1984, vol. 233, pp. 525-533, 1986 (Printed in Great Britain).
Analytical Biochemistry vol. 153, pp. 330-335 (1986)--"Phase Separation of Rat Intestinal Brush Border Membrane Proteins Using Triton X-114".

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